novel enzyme X, and you want to characterize this new enzyme. You measure the ve
ID: 858402 • Letter: N
Question
novel enzyme X, and you want to characterize this new enzyme. You measure the velocity of the reaction with different substrate concentrations and get the following data:
a) Graph the above data. Determine Km and Vmax.
Please be sure to show all excel data tables and graphs used. Please clearly show all steps and equations.
b) Is X an allosteric enzyme? Explain.
c) You decide to do this experiment again, but this time with only one third of the enzyme X concentration used in the first experiment. Estimate Km and Vmax values and explain your reasoning.
Explanation / Answer
Km = (approximately) 10mM
b) Calculate Vmax. Show any equations and calculations.
Using the equation:
Vmax= (approx) 40 mmol/min
c) Is X an allosteric enzyme? Explain.
No, X is not allosteric.
Allosteric enzymes do not follow Michaelis-Menton kinetics.
Allosteric enzymes would give a sigmoidal curve when plotting Vi vs. [S]
d) You decide to do this experiment again, but this time with only one third of the enzyme X concentration used in the first experiment. Draw a new graph on the same graph that you did the first graph on. Estimate Km and Vmax from the new graph.
Km stays the same as in part a. Vmax = 1/3 original Vmax = (approx) 13
e) You wish to find the amino acid sequence of the enzyme X. What methods might you use to determine this? Name at least three.
Acid hydrolysis, base hydrolysis, enzyme digestion (exopeptidase/endopeptidase), and Edman degradation.
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